Regulation of glutamatergic signalings by niacrophage/microglia

噬菌体/小胶质细胞对谷氨酸信号的调节

基本信息

批准号：
14571764
负责人：
NAKANISHI Hiroshi
金额：
$ 1.86万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571764/
关键词：
macrophage microglia NMDA receptor electrophysiology conditioned medium 電気生理

项目摘要

To elucidate the influence of macrophage/microglia on glutamatergic synaptic transmission in the brain and bone tissues, we first examined the effects of primary cultured microglia transferred onto the organotypic cortical slice cultures. In these microglia-transferred cortical slice cultures, stimulation of the subcortlcal white matter induced fast excitatory postsynaptic potentials followed by N-merhyl-D-aspartate (NMDA) receptor-mediated plateau-like potentials that were never observed in control slice cultures. A similar potentiation of NMDA receptor-mediated postsynaptic responses was also observed by an application of a microglial-conditioned medium (MCMI 10% v/v) in acute cortical slices. These effects of MCM disappeared after boiling or incubation with proteinase K. After fractionation of MCM by anion-exchange chromatography, the enhancing activity of each fraction was quantitated electrophysiologically. When each fraction was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the fraction 24 which showed the most potent enhancing activity on NMDA receptor-mediated responses contained a relatively strong protein band with a molecular mass of approximately 70 kDa. MCM also enhanced both glutamate-and NMDA-induced inward currents recorded from acutely isolated cortical neurons. It was also noted that glutamate and NMDA induced transient large inward currents during an application of MCMI which were never observed in the control condition. These observations strongly suggest that NMDA receptor-mediated responses were potentiated by both heat-and protease-labile (presumably 70 kD-protelns) molecules released from microglia.

为了阐明巨噬细胞/小胶质细胞对脑和骨组织中谷氨酸能突触传播的影响，我们首先研究了转移到器官皮质切片培养物中的原代培养的小胶质细胞的影响。在这些小胶质细胞转移的皮质切片培养物中，刺激皮质下白质诱导的快速兴奋性突触后电位，然后是N-甲基-D-Appartate（NMDA）受体介导的高原样潜力，在对照切片培养物中从未观察到。通过在急性皮质切片中应用小胶质细胞调节培养基（MCMI 10％V/V），也观察到了NMDA受体介导的突触后反应的类似增强。 MCM的这些作用在沸腾或与蛋白酶K孵育后消失。通过阴离子 - 交换色谱分馏MCM分馏后，从电生理学上定量了每个馏分的增强活性。当通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分析每个部分时，馏分24显示出对NMDA受体介导的反应最有效的活性，其中包含一个相对强的蛋白质带，其分子质量约为70 kDa。 MCM还增强了从急性分离的皮质神经元记录的谷氨酸和NMDA诱导的内电流。还注意到，在使用MCMI期间，谷氨酸和NMDA在对照条件下从未观察到谷氨酸诱导的瞬态大内向电流。这些观察结果强烈表明，NMDA受体介导的反应均通过从小胶质细胞释放的热和蛋白酶 - 蛋白酶 - 比脂（大概是70 kD-protelns）分子增强。